ELISpot Flex: Human IgG (ALP)

ELISpot Flex: Human IgG (ALP)

3850-2A

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Our most adaptable ELISpot format, made to fit into your unique plan. Combine it with a substrate, plate, and/or antigen of choice.

Components

Capture mAbMT91/145
Detection mAbMT78/145, biotin
Enzyme conjugateStreptavidin-ALP
StimuliR848
Recombinant human IL-2

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ELISpot plates, white


Product specifications

Intended use

This ELISpot Flex kit is intended for the enumeration of B cells secreting human IgG using the ELISpot assay. B cells secreting antigen-specific IgG, as well as B cells secreting IgG irrespective of antigen specificity (total IgG) can be enumerated. For research use only. Not for use in diagnostic procedures.

Serum/Plasma samples

Antigen-specific B cells can be enumerated in two alternative ways; by coating with antigen or by using biotinylated antigen in the detection step. For memory B cells, we recommend pre-stimulation with R848 and recombinant human IL-2 (both included).

Recommendation

Product details

ProductELISpot Flex: Human IgG (ALP)
ApplicationELISpot
AnalyteIgG
ReactivityHuman, Non-human primates
Specificity

This kit is based on a matched pair of mAbs specific for the Fc part of human IgG.

Cross-reactivity

The mAbs react with human IgG and cross-react with IgG from non-human primates (NHP). For more information please check the Cross-reactivity guide section below.

Shipping and Storage

Shipping

Shipped at ambient temperature.

Storage

Store antibodies and enzyme conjugate at 4-8 °C upon receipt. R848 and IL-2 should be stored at -20 °C.

Shelf lifeAt least 18 months from date of receipt.
PBMC were pre-incubated for 72 h in the absence or presence of R848 and recombinant IL-2. The cells were then washed, added into ELISpot wells and analyzed for the secretion of total IgG (50,000 cells/well, overnight incubation).
Datasheet/Protocol
SDS

Learn how to combine Mabtech’s antibodies for human IgG ELISA and ELISpot. Adjust the setup to detect either total IgG or antigen-specific IgG. Different antibody combinations of monoclonal antibodies have been selected for the detection of the human IgG subclasses in ELISA and ELISpot. There is no cross-reactivity to human IgA1, IgA2, IgM, and IgE.

 

Application guide: ELISA

The human IgG ELISA can be adapted to detect antigen-specific IgG or the total IgG in serum or plasma samples.

Antigen-specific IgG

Capture: Antigen
Detection mAbs: MT78/145, biotin or MT78, ALP

elisa igg 1

Total IgG

Capture mAb: MT145, unconjugated
Detection mAb: MT78, ALP

elisa igg 2

 

Application guide: ELISpot

The human IgG ELISpot and FluoroSpot can be adapted to detect antigen-specific IgG-secreting cells or the total IgG-secreting cells in cell samples like PBMCs.

Antigen-specific IgG

Capture:
Antigen
Detection mAbs:
MT78/145, biotin

elispot human igg 1

 

Capture mAbs:
MT91/145, unconjugated
Detection:
Antigen-biotin

elispot human igg 2

Total IgG

Capture mAbs:
MT91/145, unconjugated
Detection mAbs:
MT78/145, biotin

elispot human igg 3

 

 

Get inspired!

Make sure to check out how researchers have used Mabtech's human IgG ELISA, ELISpot, and FluoroSpot kits in everything from vaccine development to SARS-CoV-2 research. You can find it all in our Publication database!

The systems reactive with NHPs are either based on cross-reactive human kits or specifically developed monkey kits (NHP).

Cross-reactivity verification tests have been performed in ELISpot and/or ELISA by Mabtech and/or by others. Evaluations of assays with less solid evidence of cross-reactivity are shown as symbols within parentheses.

 Rhesus MacaqueCynomolgus MacaquePig-tailed MacaqueBaboon*Sooty MangabeyAfrican Green MonkeyNight Monkey*Common MarmosetSquirrel Monkey
ASSAYOld worldNew world
ApoA1      
ApoB       
ApoE (NHP)       
ApoE       
ApoH       
CCL2 (MCP-1)       
CCL4 (MIP-1β) ( ) ( )( )( )( )
CCL22 (MDC) ( ) ( )( )( )( )
CD25       
GM-CSF( )( )   
Granzyme A        
Granzyme B (NHP)       
Granzyme B     
IFN-α2        
IFN-α pan   ( )   
IFN-γ (NHP)
IFN-γ( )
IgA (NHP)       
IgA       
IgG       
IgM       
INS     
IL-1α        
IL-1β        
IL-2 (NHP)
IL-3       
IL-4( )( )( )( )
IL-5( )( )( ) 
IL-6( )( ) ( ) 
IL-8 (NHP)        
IL-8 (ELISA)        
IL-8 (ELISpot)        
IL-10 (NHP)      
IL-10       
IL-12/-23 (p40)      
IL-12 (p70)        
IL-13 (NHP)( )( )( )  
IL-13        
IL-17A (NHP)( )( )   
IL-17A     
IL-17F        
IL-17A/F        
IL-21      
IL-22        
IL-23     
IL-27        
IL-31        
IP-10       
Perforin( )   
TGF-β1 (latent)       
TNF-α (NHP)( )  ( )( )( )( )
TNF-α       
          
SINGLE mAbs         
Cell stimulation         
CD3, mAb CD3-1       
CD3, mAb CD3-2       
CD28, mAb CD28-A       
Flow cytometry          
IFN-γ mAb 1-D1K( )( )( )( )( )( )
IL-2 mAb MT8G10( )( )( )( ) ( )( )( )
IL-4 mAb IL4-3( )       
IL-17A mAb MT504        
Perforin mAb Pf-344     
TNF-α mAb MT15B15       
Ig reagents         
IgG1 mAb MTG1218       
IgG1 mAb MT1939       
IgG2 mAb H6200       
IgG2 mAb MTG211E       
IgG3 mAb MTG34       
IgG4 mAb MTG42       
Western blot         
IFN-γ mAb MT111W( )( )( )( )( )( )( )( )( )
TNF-α mAb MT15B15( )( )( )  ( )( )( )( )

Symbol keys:


Cross-reactivity

Verified by Mabtech and/or by others. Tests performed in ELISpot and/or ELISA.

  • good
  • poor
  • no

Indications of Cross-reactivity

Based on reports from others or by analysis using recombinant proteins.

  • ( ) good
  • ( ) poor
  • ( ) no

Monkey kits

Specifically developed monkey kits are marked with (NHP).

Blank boxes

Represent cross-reactivities not evaluated.

* Comprises several species. Cross-reactivity may have to be verified on a species basis.

Tutorial, Published December 28, 2023

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Cell stimulation in ELISpot and FluoroSpot
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Tutorial, Published November 21, 2022

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Tutorial, Published November 7, 2022

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What is a positive ELISpot response?
DFR is a non-parametric statistical test for determining positive responses in ELISpot and FluoroSpot.

Tutorial, Published September 2, 2022

Isolation, freezing, and thawing of PBMCs
This is a protocol on how to best isolate, freeze, and thaw PBMCs.

Highlighted research, Published December 16, 2021

SARS-CoV-2-specific immune responses in patients with autoimmune diseases
Patients with rheumatic diseases showed T cell responses comparable to healthy control. Patients with MS and after mRNA vaccination show different memory B cell IgG responses. SARS-CoV-2 specific immune responses in autoimmunity was analyzed using FluoroSpot, ELISpot Path, and Mabtech IRIS.

Highlighted research, Published December 2, 2021

Importance of studying SARS-CoV-2-specific memory B cells
When studying antibodies in circulation, remember that memory B cells might hide out of sight. In this study, detection of SARS-CoV-2-specific T and memory B cells 6 and 15 months after infection.

Highlighted research, Published May 30, 2020

B cell FluoroSpot for sensitive ASC detection
Mabtech IRIS and IgG/IgA/IgM FluoroSpot were used to study antibody-secreting plasma cells (ASCs) specific for the nucleocapsid (N) protein. FluoroSpot was shown to be more sensitive than e.g., ELISA.

Highlighted research, Published May 30, 2020

B cell FluoroSpot for sensitive ASC detection
Mabtech IRIS and IgG/IgA/IgM FluoroSpot were used to study antibody-secreting plasma cells (ASCs) specific for the nucleocapsid (N) protein. FluoroSpot was shown to be more sensitive than e.g., ELISA.

Highlighted research, Published October 15, 2019

Detection of tumor infiltrating B cells
Tumor infiltrating B cells are able to actively secrete immunoglobulins as shown by performing an IgA, IgG and IgM FluoroSpot. The FluoroSpot technology enabled investigation of the functionality of cells directly isolated from the tumor.

Highlighted research, Published August 18, 2018

A B-cell FluoroSpot assay to assess the risk of rejection
Reversed B-cell FluoroSpot enables the simultaneous study of multiple antigen specificities in the one well. THe assay has been used for Malaria and Dengue virus research, and now for the first time in the context of organ transplantation.
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IgG

Analyte description

Immunoglobulin G (IgG) is the most abundant Ig isotype in serum, making up approximately 80% of all serum immunoglobulins. In humans, there are four subclasses of immunoglobulin G, with the highest serum concentrations of IgG1 followed by IgG2, IgG3, and IgG4. In mice, the IgG subclasses are defined as IgG1, IgG2a/c, IgG2b, and IgG3. The IgG molecule consists of two heavy and two light chains (κ or λ), resulting in a molecule with two arms for antigen binding. High levels of IgG antibodies are induced following the initial IgM response in a typical immune response to antigens.

Alternative namesImmunoglobulin G, IgG, IgG3
Cell typeB cell
Gene ID3500, 3501, 3502